【摘要】 目的探讨通过RNAi下调非小细胞肺癌局部粘着斑激酶(FAK)的活性后对吉西他滨药理的影响。方法靶向FAK的shRNA重组质粒转染并下调细胞FAK蛋白表达,用Western Blot检测转染细胞中FAK的下调;利用MTT检测吉西他滨在不同浓度下对转染细胞增殖的影响;用流式细胞术、PI荧光染色检测吉西他滨作用下转染细胞的凋亡;Caspase与Akt活性分别用Apo-ONETM均质Caspase-3/7检测系统、Western blot检测。结果在吉西他滨不参与的情况下,FAK RNAi不能影响细胞的增殖和凋亡,但FAK RNAi明显增加了吉西他滨对肿瘤细胞的灵敏度,Akt活性的下调与这一现象有关。结论靶向FAK的shRNA重组质粒下调细胞FAK蛋白表达能够增加吉西他滨对非小细胞肺癌的细胞毒性。更多还原

【Abstract】 OBJECTIVE The hypothesis Focal adhesion kinase(FAK) of Gemcitabine chemoresistance in non-small cell lung cancer cells and examined the effect of inhibiting FAK expression on Gemcitabine-induced cytotoxicity in vitro was determined.METHODS Expression of FAK was suppressed using a plasmid encoding hairpin RNA targeting human FAK mRNA.FAK expression was quantified by Western blots.Gemcitabine-induced cytotoxicity was quantified by MTT and apoptosis was characterized by PI staining and flow cytometry.Both Caspase and Akt activity was determined by Apo-ONE Homogeneous Caspase-3/7 assay system and Western blot respectively.RESULTS While not affecting cellular proliferation or apoptosis in the absence of Gemcitabine,FAK RNAi potentiated Gemcitabine-induced cytotoxicity in vitro.FAK RNAi treatment suppressed Akt activity,which might contribute to its chemosensitizing effect.CONCLUSION Suppression of FAK expression by a plasmid encoding hairpin RNA increases Gemcitabine cytotoxicity in non-small cell lung cancer.更多还原